GSM7049824: Dex; Homo sapiens; RNA-Seq
Total RNA from the cells was isolated using the Trizol (Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions, followed by a DNase I digestion for 30 minutes to remove genomic DNA Total RNA was treated with the RiboMinus kit (Life Technologies) to remove the ribosomal RNAs. The remaining RNAs were fragmented and reverse-transcribed using random primers and reverse transcriptase. Second strand cDNA synthesis was completed using DNA polymerase I and RNase H. The Illumina series KAPA Library Preparation Kit (Kapa Biosystems) was used to perform end repair, dA-tailing, and adapter ligation. The dUTP-marked second strand was digested at 37 °C for 30 min using Uracil Specific Excision Reagent (USER) enzyme (New England Biolabs). The cDNA was enriched by PCR amplification to create the final cDNA libraries.